Department of Biomedical Engineering, National Cheng Kung University, No. 1 University Rd., Tainan, Taiwan.
Biomed Microdevices. 2013 Feb;15(1):135-43. doi: 10.1007/s10544-012-9695-y.
Microelectrode arrays (MEAs) incorporated with the electric cell substrate impedance sensing (ECIS) technique provide a method for acquiring cellular electrophysiological information, which is useful for the time-course monitoring of cellular outgrowth and damage. This research utilizes the ECIS technique for monitoring the time-course impedimetric changes in normal and insulin-like growth factor 1 (IGF-1)-protected cortical neurons under the ischemic insult of oxygen glucose deprivation (OGD) created in a microperfusion environment. The neuronal apoptosis is reflected by the relatively low cell viability (28 ± 11.5 %) after 30-min OGD followed by 24 h of re-oxygenation. Also the hyperpolarization phase of mitochondrial membrane potential (MMP) occurs during 2 h of the re-oxygenation period. In contrast, cortical neurons treated with 50 and 100 ng/mL IGF-1 show higher survival rates of 45 ± 5.2 % and 49 ± 9.2 %, respectively, and no occurrence of the hyperpolarization of MMP during the re-oxygenation period. The ECIS results demonstrate that the measured impedance of cortical neurons decreased from 826 ± 86 kΩ to 224 ± 32 kΩ due to cell detachment under the insult of OGD. The measured impedance of IGF-1-protected cortical neurons slowly decreased to about 50 % of the original value (560 ± 45 kΩ for 50 ng/mL and 593 ± 44 kΩ for 100 ng/mL) compared to saline control of 232 ± 37 kΩ, which indicates improved cell adhesion under OGD conditions. The time-course impedimetric results show that the proposed ECIS-based MEAs platform incorporated with a microperfusion environment can be used for the real-time monitoring of cortical neurons under in vitro OGD and the IGF-1 protective effect against OGD-induced ischemic neuronal death.
微电极阵列 (MEA) 与电细胞基质阻抗传感 (ECIS) 技术相结合,提供了一种获取细胞电生理信息的方法,可用于细胞生长和损伤的时间过程监测。本研究利用 ECIS 技术监测在微灌注环境中创建的氧葡萄糖剥夺 (OGD) 缺血应激下正常和胰岛素样生长因子 1 (IGF-1) 保护的皮质神经元的时间过程阻抗变化。神经元凋亡通过 30 分钟 OGD 后 24 小时再氧合后的相对低细胞活力(28±11.5%)反映。此外,线粒体膜电位(MMP)的超极化阶段发生在再氧合期的 2 小时内。相比之下,用 50 和 100ng/mL IGF-1 处理的皮质神经元显示出更高的存活率,分别为 45±5.2%和 49±9.2%,并且在再氧合期间没有发生 MMP 的超极化。ECIS 结果表明,由于 OGD 损伤导致细胞脱落,皮质神经元的测量阻抗从 826±86kΩ 降低到 224±32kΩ。与生理盐水对照(232±37kΩ)相比,IGF-1 保护的皮质神经元的测量阻抗缓慢降低至原始值的约 50%(50ng/mL 时为 560±45kΩ,100ng/mL 时为 593±44kΩ),这表明在 OGD 条件下改善了细胞黏附。时间过程阻抗结果表明,所提出的基于 ECIS 的 MEA 平台与微灌注环境相结合,可用于体外 OGD 下皮质神经元的实时监测以及 IGF-1 对 OGD 诱导的缺血性神经元死亡的保护作用。